菜心叶片响应高温胁迫的转录组分析及转录因子筛选
Transcriptome Analysis of Chinese Flowering Cabbage Leaves and Tran-scription Factor Screening under Heat Stress
庞强强 1孙晓东 1周曼 1蔡兴来 1陈贻诵 1王亚强1
作者信息
- 1. 海南省农业科学院蔬菜研究所,海南省蔬菜学院士团队创新中心,海南省瓜菜育种工程技术研究中心,海南省蔬菜生物学重点实验室,海口,571100
- 折叠
摘要
为探索菜心响应高温胁迫的分子机制,挖掘高温胁迫相关基因.利用转录组测序技术,对热敏DH系CX7-3和耐热DH系CX1-7高温胁迫处理前后叶片中的转录情况进行分析,建立差异表达基因库,并从中筛选高温胁迫转录因子.结果表明,转录组测序共获得32.25 Gb的样本数据和46 204条unigene,在9个功能数据库中有76.76％的unigene得到注释.比较高温胁迫前后样本,获得3 803个差异表达基因.GO功能分类可将DEG富集到生物过程、细胞组分和分子功能3个方面,KEGG功能富集数量居于前三位的为植物-病原相互作用、植物激素信号转导和植物MAPK信号通路.从差异表达基因库中鉴定得到275个转录因子,主要涉及MYB、AP2、C3H、NAC、C2H2、bHLH、bZIP、HSF和WRKY等34类转录因子家族,且受高温诱导呈现多种表达模式.进一步筛选获得15个共表达、4个特有表达、1个非特有表达转录因子基因.挑选12个转录因子基因进行RT-qPCR验证,基因表达趋势与RNA-seq分析结果一致.本研究结果为今后开展菜心耐热分子机制和耐热新品种的选育提供理论参考.
Abstract
In order to explore the molecular mechanism and mining related genes in response to heat stress of Chinese flowering cabbage.Transcriptome sequencing(RNA-Seq)technology was used to analyze the transcrip-tion in leaves of heat-sensitive DH line CX7-3 and heat-resistant DH line CX1-7 under heat stress to establish dif-ferential expressed gene library and screen the transcription factor genes.A total of 32.25 Gb sample data were ob-tained by RNA-Seq,46 204 unigenes were obtained after assembly and de redundancy,76.76％unigenes can be annotated in nine functional databases.3 803 differential expressed gene were obtained through pairwise compari-son between samples under heat stress.Go function analysis shows that the differential expressed genes were en-riched into biological process subclasses,cell component subclasses and molecular functional subclasses.The top three metabolic pathways of KEGG functional enrichment were plant-pathogen interaction,plant hormone signal transduction and MAPK signaling pathway-plant.From the differential expressed genes library,we identified 275 transcription factors,mainly involving 34 transcription factor families such as MYB,AP2,C3H,NAC,C2H2,bHLH,bZIP,HSF and WRKY.RNA-Seq data showed that transcription factor genes produced multiple expres-sion patterns induced by high temperature,and fifteen co-expressed genes,four specific-expressed and one non specific-expressed transcription factor gene were further screened.Twelve high temperature response-related tran-scription factor genes were analyzed by RT-qPCR,the gene expression trend was consistent with the results of RNA-Seq analysis.The results of this study provide theoretical reference for the further research for the molecular mechanism to heat stress and variety breeding in the future.
关键词
菜心/高温胁迫/转录组/差异表达基因/转录因子Key words
Chinese flowering cabbage(Brassica campestris L.ssp.chinensis var.utilis Tsen et Lee)/Heat stress/Transcriptome/Differential expressed gene/Transcription factor引用本文复制引用
基金项目
海南省科技专项-省属科研院所技术创新科研项目(SQKY2022-0011)
国家现代农业产业技术体系建设专项(CARS-23-G52)
海南省自然科学基金项目(318QN293)
出版年
2024
NETL
NSTL
万方数据